Chiral separation of new sulfonamide derivatives and evaluation of their enantioselective affinity for human carbonic anhydrase II by microscale thermophoresis and surface plasmon resonance.

摘要: Abstract The aim of this study was to develop a method combining chiral separation and biophysical techniques evaluate the enantioselective affinity original sulfonamide derivatives towards their therapeutic target, human carbonic anhydrase II (hACII). first step consisted in preparation enantiomers by chromatographic separation. performances HPLC Supercritical Fluid Chromatography (SFC) were studied at analytical scale optimization various experimental conditions using adsorbed polysaccharide stationary phases (amylose AD-H cellulose OD-H). Since SFC allowed obtaining higher enantioresolutions per time unit, it selected for semi-preparative successfully used isolate each enantiomer with satisfactory enantiomeric purity (>98%). Secondly, microscale thermophoresis (MST) surface plasmon resonance (SPR) as reference developed measure potential affinities these hACII. optimizations both methods performed compound, i.e. acetazolamide, which hCAII has previously been demonstrated. For all compounds, K D values obtained MST SPR good agreement, leading similar scales despite approaches totally differ (labeling versus immobilization protein SPR). equilibrium dissociation constants our compounds range 100–1000 nM an enantioselectivity observed diarylpyrazole 2 . Finally, comparing techniques, appears especially adapted further screening series due lower required estimate binding constant while consuming little SPR.